These 5172 3-state Cre-sensitive (5172 3SCS) mice contain a construct designed to insert (from 5' to 3') a floxed-STOP cassette, tdTomato open reading frame (ORF), and a floxed-internal ribosome entry site (IRES) fused to an enhanced green fluorescent protein (GFP) ORF. The IRES-eGFP element was flanked by 5172 mutant loxP sites (which recombine at 30% the efficiency of wildtype loxP sites). This vector was inserted into the Gt(ROSA)26Sor locus allowing for widespread expression. These 5172 3SCS mice are of general use for detecting cells that have expressed the Cre recombinase, for Cre recombinase-based lineage marking experiments, and in particular for applications in which it is useful to discriminate cells that differ in how much Cre recombinase they have expressed.
Cre recombinase-dependent excision of IRES-GFP in the 5172 3SCS strain occurs at approximately three times the efficiency it does in the 3373 3SCS strain (Stock No. 013586).
Nigel Killeen, Univ of California San Francisco (UCSF)
These 5172 3-state Cre-sensitive (5172 3SCS) mice contain a construct designed to insert (from 5' to 3') a floxed-STOP cassette, tdTomato open reading frame (ORF), and a floxed-internal ribosome entry site (IRES) fused to an enhanced green fluorescent protein (GFP) ORF. The IRES-eGFP element was flanked by 5172 mutant loxP sites (which recombine at 30% the efficiency of wildtype loxP sites). This vector was inserted into the Gt(ROSA)26Sor locus allowing for widespread expression. The floxed-STOP cassette causes termination of transcription and results in no expression of either fluorophore. When bred to mice that express Cre recombinase, offspring will have the floxed-STOP cassette deleted in the cre-expressing tissue(s). This deletion results in tdTomato and/or GFP expression depending on how much Cre-recombinase the cells express. When crossed with a strain expressing low amounts of Cre recombinase, partial recombination results in tdTomato+ GFP+ cells due to removal of the floxed-STOP. When crossed to strains with higher concentrations of cre, the 5172 mutant loxP sites flanking the IRES-GFP cassette will be recombined and will result in tdTomato+ cells with varying amounts of GFP expression due to removal of the IRES-GFP cassette. These 5172 3SCS mice are of general use for detecting cells that have expressed the Cre recombinase, for Cre recombinase-based lineage marking experiments, and in particular for applications in which it is useful to
discriminate cells that differ in how much Cre recombinase they have
Cre recombinase-dependent excision of IRES-GFP in the 5172 3SCS strain
occurs at approximately three times the efficiency it does in the 3373 3SCS
strain (Stock No. 013586)
The 5172 reporter construct was designed to insert (from 5' to 3') a splice acceptor (SA) sequence, a loxP site, a puromycin selection cassette, an SV40 polyadenylation (polyA) cassette, another loxP site, tdTomato open reading frame (ORF), a loxP-flanked internal ribosome entry site (IRES) fused to an enhanced green fluorescent protein (GFP) ORF, followed by a second polyA cassette. The IRES-eGFP-polyA element was flanked by 5172 mutant loxP sites (which are recombined at 30% the efficiency of wildtype loxP sites). The loxP sites flanking the 5' polyA cassette are in opposite orientation to the loxP sites flanking the 3' IRES-GFP cassette. This vector is driven by the Gt(ROSA)26Sor promoter and was electroporated into 129P2/OlaHsd-derived E14Tg2A.4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6x(C57BL/6xDBA/2 F1) recipient blastocysts, and chimeric males were bred with C57BL/6 females. These 5172 3-state Cre-sensitive (5172 3SCS) mice were backcrossed to C57BL/6J mice for at least 5 generations. Upon arrival at The Jackson Laboratory, mutant mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.
|Allele Name||targeted mutation 3, Nigel Killeen|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||targeted mutation 3, Nigel Killeen; Gt(ROSA)26Sortm3Nik|
|Gene Symbol and Name||Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano|
|Gene Synonym(s)||beta geo; ROSA26; R26; Gtrgeo26; Gtrosa26; Gtrgeo26; Gtrosa26; AV258896; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; SETD5-AS1; Thumpd3as1|
|Site of Expression||Cre excision of the stop signal results in expression of tdTomato and/or GFP in cre-expressing tissues depending on how much Cre-recombinase the cells express.|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||The construct inserted into the locus contained (5' to 3') a splice acceptor, floxed stop cassette, tdTomato sequence, and an IRES-EGFP flanked by 5172 loxP sites.|
When maintaining a live colony, homozygotes may be bred together.
When using the B6.129P2-Gt(ROSA)26Sortm3Nik/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #013587 in your Materials and Methods section.
|Heterozygous or wildtype for Gt(ROSA)26Sor<tm3Nik>|
We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.
Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.
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